Geranyl compounds

ABSTRACT

This invention provides compounds having excellent antitumor activity, which are represented by the following formulae 
     
       
         
         
             
             
         
       
         
         
           
             in which R 1 , R 2 , R 3 , m, n and R 4  have the significations as given in the specification.

This application is a divisional of U.S. application Ser. No.11/356,019, filed Feb. 17, 2006, now U.S. Pat. No. 7,507,765, which is adivisional of U.S. application Ser. No. 10/498,882, filed Jun. 16, 2004,now U.S. Pat. No. 7,125,852, which is a national stage application ofInternational Application No. PCT/JP02/13615, filed Dec. 26, 2002.

TECHNICAL FIELD

This invention relates to novel geranyl compounds or mevalonic acidderivatives, and to their utilization as antitumor agents.

BACKGROUND ART

Many geranyl compounds having 1,5-diene structure are present in vivo,and are known as in vivo precursors of substances having polyenestructure and exhibiting various physiological activities. Thesesubstances having 1,5-diene structure and polyenes derived therefrominvariably start from mevalonic acid and biosynthesized.

I noticed, as such geranyl compounds having 1,5-diene structure,germanic acid or geranylamine, and furthermore mevalonic acid which isthe base for biosynthesis of polyenes, synthesized various derivativesof germanic acid or geranylamine and mevalonic acid and investigatedtheir physiological activities, in particular, antitumor activity andtoxicity, and have come to complete the present invention.

DISCLOSURE OF THE INVENTION

This invention provides geranyl compounds represented by the followingformulae (I-1), (I-2) or (I-3):

in which

R¹ stands for

R² stands for a residual group remaining after removing all carboxylgroups present in a carboxylic acid selected from the group consistingof malic acid, citric acid, succinic acid, fumaric acid, 2-oxoglutaricacid, pyruvic acid, p-pyruvoaminobenzoic acid, retinoic acid, tyrosine,cysteine, glutamic acid and serine, and where hydroxyl or amino group(s)are present in the residual group, they may optionally be protected byacyl (e.g., lower alkanoyl) or benzyloxycarbonyl group(s),

m is 1, 2 or 3,

n is 0, 1 or 2,

m+n showing the number of carboxyl groups which are present in saidcarboxylic acid, and

R³ stands for p-hydroxyphenyl or mercapto group.

The invention also provides mevalonic acid derivatives represented bythe following formula (I-4):

in which R⁴ stands for —CH₂OH or —CH₃.

Those geranyl-sugar derivatives of above formula (I-1) include thefollowing five compounds:

N-geranylglucuronamide

N-geranylgalacturonamide

N-galactosylgeranamide

N-glucosylgeranamide

and

N-fucosegeranamide

The geranylamide derivatives of above formula (I-2) include, forexample, the following compounds.

N,N′-digeranylmalic Diamide

O-acetyl-N-geranylmalic Monoamide

O-acetyl-N,N′-digeranylmalic Diamide

N,N′,N″-trigeranylcitric Triamide

N-geranylsuccinic Monoamide

N,N′-digeranylsuccinic Diamide

N,N′-digeranylfumaric Diamide

N-geranylfumaric Monoamide

N,N′-digeranyl-2-oxoglutaric Diamide

N-geranylpyruvamide

N-geranyl-p-pyruvoaminobenzamide

Tyrosine Geranylamide

N-acetyltyrosine Geranylamide

Cysteine Geranylamide

Glutamic Digeranyldiamide

Serine Geranylamide

and

N-geranyl Retinamide

Also the geranylamide derivatives of above formula (I-3) include thefollowing two compounds:

N-geranoyltyrosine

and

N-geranoylcysteine

The mevalonic acid derivatives of above formula (I-4) include, forexample, the following:

N-glucosylmevalonamide

N-galactosylmevalonamide

and

N-fucosemevalonamide

Among the compounds of the formula (I-1), those of the formulae (1) and(2) can be prepared by, for example, subjecting geranylamine to anamidation reaction with reactive derivatives (e.g., mixed acidanhydride, active ester, halide or the like) of glucuronic acid orgalacturonic acid whose hydroxyl group(s) are protected with acylgroup(s) (e.g., acetyl).

Of the compounds of the formula (I-1), those of the formulae (3) to (5)can be prepared by, for example, subjecting reactive derivatives ofgeranic acid (e.g., mixed acid anhydride, active ester, halide or thelike) to an amidation reaction with galactosamine, glucosamine orfucosamine.

Said amidation reaction can be conducted following the conventionalmethod of amidation reaction in the field of peptide chemistry, normallyin an adequate inert organic solvent (e.g., tetrahydrofuran, chloroform,N,N-dimethylformamide, dichloromethane or the like) or in water, undercooling down to about 0° C. or heating up to about 60° C., preferably atabout 0° C. to room temperature.

The use ratio of geranylamine to a reactive derivative of gulcuronicacid or galacturonic acid whose hydroxyl group(s) are protected is notstrictly limited, but it is normally preferred to use the geranylaminewithin a range of 1-2 moles, per mole of the reactive derivative.

The use ratio of galactosamine, glucosamine or fucosamine to a reactivederivative of geranic acid is again not strictly limited, but normallyit is preferred to use galactosamine, glucosamine or fucosamine within arange of 1-2 moles, per mole of the reactive derivative.

Where hydroxyl-protective groups are present after the amidationreaction, said protective groups are removed by a de-protection reactionsuch as hydrolysis, to provide geranyl-sugar derivatives of the formula(I-1).

The geranyl-sugar derivatives of the formula (I-1) produced throughabove reactions can be isolated from the reaction mixtures and purifiedby conventional means, for example, extraction, crystallization,chromatography or the like.

The geranylamide derivatives of the above formula (I-2) can be producedby, for example, subjecting geranylamine to an amidation reaction withreactive derivatives (e.g., mixed acid anhydride, active ester, halideor the like) of carboxylic acid represented by the formula (II):

in which R², m and n have the earlier given significations, in whichhydroxyl or amino group(s) are protected with acyl (e.g., lower alkanoylsuch as acetyl), benzyloxycarbonyl and the like groups.

Said amidation reaction can be conducted following the conventionalmethod of amidation reaction in the field of peptide chemistry, normallyin an adequate inert organic solvent (e.g., tetrahydrofuran, ether,dichloromethane, chloroform, N,N-dimethylformamide or the like) undercooling down to about 0° C. or heating up to about 60° C., preferably atabout 0° C. to room temperature.

The use ratio of geranylamine to a reactive derivative of carboxylicacid of the formula (II) is variable depending on the number of geranylgroup (m) to be introduced into the carboxylic acid, while it isnormally preferred to use it within a range of 1 mole to (m+2) moles permole of the reactive derivative.

When the hydroxyl- or amino-protective groups are present after theamidation reaction, they are removed where necessary by a de-protectionreaction such as hydrolysis to provide geranyl amide derivatives of theformula (I-2).

Those geranylamide derivatives of the formula (I-3) can be prepared by,for example, subjecting reactive derivatives of geranic acid (e.g.,mixed acid anhydride, active ester, halide and the like) to an amidationreaction with tyrosine or cysteine.

This amidation reaction can also be conducted following conventionalmethod of amidation reaction in the field of peptide chemistry, normallyin an adequate inert organic solvent (e.g., tetrahydrofuran, ether,dichloromethane, chloroform, N,N-dimethylformamide or the like) or inwater, under cooling down to about 0° C. or heating up to about 60° C.,preferably at about 0° C. to room temperature.

The use ratio of tyrosine or cysteine to a reactive derivative ofgeranic acid is not strictly limited, but it is normally preferred touse either of them within a range of 1-2 moles per mole of the reactivederivative.

Such geranylamide derivatives of the formula (I-2) or (1-3) produced inthe above reactions can be isolated from the reaction mixtures andpurified by conventional means, for example, extraction,crystallization, chromatography or the like.

Those mevalonic acid derivatives of the formula (I-4) can be prepared,for example, by reacting sugaramine represented by the followingformula:

in which R⁴ has the earlier given signification, or salt thereof withmevalolactone or mevaloyl halide.

Said reaction of a sugaramine of the formula (III) or a salt thereofwith mevalolactone or mevaloyl halide (e.g., mevaloyl chloride) can beconducted in water or an adequate inert organic solvent (e.g.,N,N-dimethylformamide, tetrahydrofuran, chloroform or the like) attemperatures between room temperature to reflux temperature of thesolvent, preferably from about 40° to about 70° C.

The use ratio of mevalolactone or mevaloyl halide to a sugaramine of theformula (III) is not strictly limited, but it is normally preferred touse 1-2 moles of mevalolactone or mevaloyl halide per mole of thesugaramine of the formula (III).

Where a salt of a sugaramine of the formula (III) or mevaloyl halide isused as the starting material, it is generally desirable to carry outthe above reaction with addition of a base, for example, tertiary aminesuch as N-methylpiperidine; or an inorganic base such as sodiumhydroxide, potassium hydroxide, potassium carbonate and the like.

The mevalonic acid derivatives of the formula (I-4) produced throughabove reactions can be isolated from the reaction mixtures and purifiedby conventional means, such as extraction, crystallization,chromatography or the like.

Compounds of the formulae (I-1) through (I-4) offered by the presentinvention possess excellent antitumor activity, as is clear from thefollowing measurement results of antitumor effect.

Measurement of Antitumor Effect

Carcinoma of HuH-7 cells (dendriform cell strain of human hepatoma)hypodermically implanted or subimplanted in the backs of 5-weeks oldfemale nude mice (BALB/c, Ninox) was aseptically taken out and crushedinto 5×5 mm sized pieces in a phosphate buffer solution (PBS), a pieceof which then being hypodermically implanted in the backs of the nudemice.

Each test substance was dissolved in corn oil, and the solution wasintraperitoneally administered to the nude mice consecutively once perday at a rate of 250 μg/mouse for 3 weeks, starting from a week afterthe implantation. After termination of the administration, thecarcinomas were taken out and weighed to calculate the antitumor effectand the mice' weight loss by the following equations. For the test sixmice per group were used, and the group administered with the solvent(corn oil) alone was made the control group:

${{Antitumor}\mspace{14mu}{effect}\mspace{14mu}(\%)} = {\frac{{average}\mspace{14mu}{tumor}\mspace{14mu}{weight}\mspace{14mu}{of}\mspace{14mu} a\mspace{14mu}{test}\mspace{14mu}{group}}{{average}\mspace{14mu}{tumor}\mspace{14mu}{weight}\mspace{14mu}{of}\mspace{14mu}{the}\mspace{14mu}{control}\mspace{14mu}{group}} \times 100}$${{Weight}\mspace{14mu}{loss}\mspace{14mu}(\%)} = {\frac{{average}\mspace{14mu}{body}\mspace{14mu}{weight}\mspace{14mu}{of}\mspace{14mu} a\mspace{14mu}{test}\mspace{14mu}{group}\mspace{14mu}{mice}}{{average}\mspace{14mu}{body}\mspace{14mu}{weight}\mspace{14mu}{of}\mspace{14mu}{the}\mspace{14mu}{control}\mspace{14mu}{group}\mspace{14mu}{mice}} \times 100}$

Evaluations of antitumor effect and weight loss were conducted accordingto the following standard, where the control group values were held tobe 100%.

Antitumor Effect

-   -   −: >100%, +/−: 100˜75%, +: 75˜50%, ++: 50˜25%, +++: 25˜0%        Weight Loss    -   −: >110%, +/−: 110˜100%, +: 100˜95%, ++: 95˜90%, +++: <90%        Death Rate (Death Rate During the Test Period)    -   −: none    -   +/−: death occurred with high concentration administration (500        μg/mouse)    -   +: 1-3 mice dead    -   ++: 3-5 mice dead    -   +++: all 6 mice dead        Synthetic Evaluation    -   −: weak antitumor effect and very strong toxicity against host        mice    -   +/−: weak antitumor effect recognizable and toxicity against        host mice also observable    -   +: a fixed level of antitumor effect observable but toxicity        against host mice also strong    -   ++: strong antitumor effect observed and toxicity against host        mice week    -   +++: strong antitumor effect observed and no toxicity against        host mice

The results are shown in the following Tables 1-3.

TABLE 1 Toxicity Antitumor Weight Death Synthetic Test Substance Effectloss rate Evaluation N-geranyl gulonamide ++ +/− − ++ N-geranyl +++ +/−+/− galacturonamide N-galactosylgeranamide +++ + − +++N-fucosegeranamide ++ +/− − +++

TABLE 2 Toxicity Antitumor Weight Death Synthetic Test Substance Effectloss rate Evaluation N,N′-digeranylmalic diamide + + − ++N,N′-digeranylfumaric diamide ++ +/− − ++ N-geranyl-4- ++ − − +++pyruvoaminobenzamide N-geranoyltyrosine ++ +/− − ++ Tyrosinegeranylamide + +/− − + N-acetyltyrosine geranylamide + +/− − +

TABLE 3 Toxicity Antitumor Weight Death Synthetic Test Substance Effectloss rate Evaluation N-glucosylmevalonamide ++ +/− − +++

As is clear from the shown results, the compounds of the invention ofthe formulae (I-1) to (I-4) possess excellent antitumor effect againstHuH-7 cells and furthermore almost no toxicity, and are expected to beuseful as antitumor agents for treatment and therapy of various solidcancer represented by liver cancer.

Where a compound of the present invention is used as a medicine such asan antitumor agent, it can be administered orally or parenterally (e.g.,intravenous injection, intramuscular injection, hypodermic injection, orthe like). The effective dose is variable over a broad range dependingon individual patients' symptoms, seriousness of the illness, bodyweight, age, and the doctor's diagnosis, etc. Normally, however, takinga case of administration by injection, the dose can be about 1-about 50mg/kg/day, which may be administered at a single time or at plural timesdividedly in a day.

Where a compound of the present invention is used as a medicine, aneffective dose of the compound can be formulated with pharmaceuticallyacceptable carrier or diluent (e.g., excipient, solvent or otheradjuvants) into a preparation form suitable for unit doseadministration, for example, tablet, powder, granule, capsule, entericcoated pill, troche, syrup, elixer, liquid, suspension, emulsion and thelike.

As carriers or diluents useful for the formulation, for example,excipients such as starch, lactose, sucrose, mannitol, carboxymethylcellulose and the like; lubricants such as magnesium stearate, sodiumlaurylsulfate, talc and the like; binders such as dextrin,microcrystalline cellulose, polyvinylpyrrolidone, gum Arabic, cornstarch, gelatin and the like; disintegrators such as potato starch,carboxymethyl cellulose and the like; and diluent solvents such as waterfor injection, physiological saline, aqueous dextrose solution,vegetable oil for injection, propylene glycol, polyethylene glycol andthe like can be named. Furthermore, taste- or odor-correcting agent,colorant, isotonicity, stabilizer, antiseptic, soothing agent and thelike may be incorporated where necessary.

In the pharmaceutical preparations according to the invention, moreover,other pharmacologically active substance(s) may be incorporated wherenecessary.

Hereinafter the invention is explained still more specifically,referring to working Examples.

EXAMPLES Synthesis Example 1 Synthesis of N-geranylgalacturonamide

To a tetrahydrofuran (THF) (20 ml) solution containingO-tetraacetylgalacturonic acid (3.62 g, 10 mmols), triethylamine (1.01g, 10 mmols) was added, and the solution was cooled to 0° C. Into thissolution isobutyl chloroformate (1.37 g, 10 mmols) solution in THF (5ml) was added dropwise at 0° C., followed by 30 minutes' stirring. Intothe resulting solution a geranylamine (1.53 g, 10 mmols) solution in THF(5 ml) was added dropwise, followed by an hour's stirring at 0° C. andfurther 4 hours' stirring at room temperature. After termination of thereaction, 150 ml of chloroform was added, and the chloroform layer waswashed three times each with 50 ml of water. The chloroform layer wasdried over magnesium sulfate, the chloroform was concentrated, and theresidue was purified on silica gel column chromatography. Consequently,3.58 g (73.5%) of N-geranyl-O-tetraacetylgalacturonamide was obtained asa viscous oily substance, from its hexane-acetone (3:1) distillate.

¹H NMR(CDCl₃) δ=1.58 (3H, s), 1.65 (3H, s), 1.68 (3H, s), 2.01 (3H, s),2.02 (3H, s), 2.05 (3H, s), 2.15 (3H, s), 2.02-2.11 (4H, m), 3.70-3.83(1H, m), 3.83-3.96 (1H, m), 5.00-5.17 (2H, m), 5.29 (1H, d, J=10.8 Hz),5.39 (1H, d, J=10.8 Hz), 6.29-6.46 (2H, m).

3.58 Grams (7 mmols) of above product was dissolved in 30 ml of ethanol,35 ml of 1N aqueous sodium hydroxide solution was added, and stirred for2 hours at room temperature. Then 35 ml of 1N hydrochloric acid wasadded to the reaction mixture and condensed under reduced pressure. Tothe residue 150 ml of ethanol was added and precipitated sodium chloridewas filtered off. The filtrate was again condensed, and the residue wasseparated on silica gel column chromatography. From the hexane-ethanol(3:1) distillate, 1.95 g of N-geranylgalacturonamide was obtained as aviscous compound. Ether was added to this product to conductcrystallization, and by suction filtration 1.03 g of crystallized titlecompound was obtained. The yield was 45%.

¹H NMR (DMSO-d6) δ=1.50 (3H, s), 1.55 (3H, s), 1.56 (3H, s), 1.86-2.04(4H, m), 3.48-3.76 (2H, m), 3.80-3.94 (2H, m), 4.07-4.84 (3H, m), 4.99(1H, d, J=9.6 Hz), 5.06 (1H, d, J=9.6 Hz).

Synthesis Example 2 Synthesis of N-geranylgulcronamide

Synthesis Example 1 was repeated except that O-tetraacetylglucuronicacid was used in place of O-tetraacetylgalacturonic acid, to provide thetitle compound.

¹H NMR (DMSO-d6) δ=1.60 (3H, s), 1.68 (3H, s), 1.73 (3H, s), 2.07-2.09(4H, m), 3.58-3.62 (2H, m), 3.81-3.95 (2H, m), 4.07-4.86 (3H, m),5.05-5.09 (1H, m), 5.36-5.40 (1H, m).

Synthesis Example 3 Synthesis of N-galactosylgeranamide

To a THF (20 ml) solution containing geranic acid (0.84 g, 5 mmols),triethylamine (0.51 g, 5 mmols) was added and cooled to 0° C., intowhich a THF (5 ml) solution containing isobutyl chloroformate (0.68 g, 5mmols) was added dropwise, followed by 30 minutes' stirring at 0° C.Then galactosamine hydrochloride (1.08 g, 5 mmols) was dissolved in 10ml of water, to which 5 ml of 1N sodium hydroxide was further added, andthe solution was added to the reaction mixture all at a time. After thefollowing stirring for an hour at 0° C. and for further 4 hours at roomtemperature, the reaction mixture was condensed under reduced pressure.To the residue 150 ml of acetone was added and precipitated sodiumchloride was filtered off. The filtrate was condensed once again and theresidue was separated on silica gel column chromatography. Consequentlythe product was obtained from the hexane-ethanol (2:1) distillate as aviscous oily substance, to which a minor amount of ether was added tocrystallize the product. Through subsequent suction filtration, 0.754 gof the title compound was obtained. The yield was 46%, and the meltingpoint was 85-87° C.

¹H NMR (DMSO-d6) δ=1.53 (3H, s), 1.57 (3H, s), 1.59 (3H, s), 1.95-2.01(4H, m), 3.63-3.79 (7H, m), 4.25-4.60 (3H, m), 4.85-4.92 (1H, m),5.00-5.08 (1H, m), 6.25-6.31 (1H, m).

Synthesis Example 4 Synthesis of N-glucosylgeranamide

Synthesis Example 3 was repeated except that glucosamine hydrochloridewas used in place of galactosamine hydrochloride, to provide the titlecompound.

¹H NMR (DMSO-d6) δ=1.54 (3H, s), 1.60 (3H, s), 2.03 (3H, s), 1.96-2.20(4H, m), 3.38-3.61 (4H, m), 4.38-4.66 (3H, m), 5.75 (1H, s), 6.34-6.39(1H, m).

Synthesis Example 5 Synthesis of N-fucosegeranamide

Synthesis Example 3 was repeated except that fucosamine hydrochloridewas used in place of galactosamine hydrochloride, to provide the titlecompound.

¹H NMR (DMSO-d6) δ=1.31 (3H, d, J=5.4 Hz), 1.53 (3H, s), 1.60 (3H, s),1.72 (3H, s), 1.97-2.05 (4H, m), 3.88-3.95 (2H, m), 4.21-4.24 (1H, m),4.44-4.46 (1H, m), 4.82-4.87 (1H, m), 5.00-5.13 (1H, m).

Synthesis Example 6 Synthesis of N,N′-digeranylfumaric Diamide

To a tetrahydrofuran (THF) (20 ml) solution containing fumaric acid(0.58 g, 5 mmols), triethylamine (1.01 g, 10 mmols) was added and thesolution was cooled to 0° C., into which a THF (5 ml) solutioncontaining isobutyl chloroformate (1.53 g, 10 mmols) was added dropwise.As the addition was continued, white precipitate started to form. After30 minutes' stirring at 0° C., a THF (5 ml) solution containinggeranylamine (1.53 g, 10 mmols) was added dropwise into the system,followed by an hour's stirring at 0° C. and further 4 hours' stirring atroom temperature. After termination of the reaction, 50 ml of water wasadded to the reaction mixture which was then extracted with chloroform.The chloroform layer was washed with water and dried over magnesiumsulfate. Filtering the magnesium sulfate off, the chloroform layer wascondensed to provide a white crystal. Recrystallizing the same fromethanol, 1.07 g of the title compound was obtained. The yield was 55%.

¹H NMR(CDCl₃) δ=1.60 (6H, s), 1.62 (6H, s), 1.68 (6H, s), 2.01-2.10 (8H,m), 3.95 (4H, t, J=9.6 Hz), 5.04-5.09 (2H, m), 5.20-5.25 (2H, m), 5.94(2H, brs), 6.90 (2H, s), 7.26 (2H, s).

Synthesis Example 7

Synthesis Example 6 was repeated except that the fumaric acid wasreplaced with corresponding carboxylic acid of the earlier given formula(II) in each run, to provide the following compounds:

N-geranylpyruvamide

¹H NMR(CDCl₃) δ=1.55 (3H, s), 1.64 (3H, s), 1.82 (3H, s), 2.00 (3H, s),1.92-2.12 (4H, m), 3.84 (2H, d, J=7.2 Hz), 4.96-5.12 (1H, m), 5.22-5.35(1H, m).

N,N′-digeranylmalic Diamide

¹H NMR(CDCl₃) δ=1.58 (6H, s), 1.64 (6H, s), 1.67 (6H, s), 1.94-2.14 (8H,m), 2.54 (1H, dd, J=4.8, 14.8 Hz), 2.79 (1H, dd, J=3.2, 14.4 Hz),3.75-3.93 (4H, m), 4.32-4.40 (1H, m), 5.00-5.10 (2H, m), 5.10-5.22 (2H,m).

O-acetyl-N-geranylmalic Monoamide

¹H NMR(CDCl₃) δ=1.60 (3H, s), 1.68 (3H, s), 1.69 (3H, s), 1.96-2.11 (4H,m), 2.19 (3H, s), 2.65 (1H, dd, J=9.6, 22.8 Hz), 3.00 (1H, dd, J=2.4,22.8 Hz) 3.79-3.89 (2H, m), 4.51-4.56 (1H, m), 5.08 (1H, t, J=7.2 Hz),5.18 (1H, t, J=6.0 Hz).

O-acetyl-N,N′-digeranylmalic diamide, which was synthesized by a similarmethod, using N-geranylmalic acid monoamide as the starting material.

¹H NMR(CDCl₃) δ=1.59 (6H, s), 1.67 (6H, s), 1.68 (6H, s), 1.94-2.01 (8H,m), 2.16 (3H, s), 2.55 (1H, dd, J=13.2, 22.8 Hz), 2.97 (1H, dd, J=2.4,22.8 Hz), 3.79-3.89 (4H, m), 4.34-4.40 (1H, m), 5.02-5.10 (2H, m),5.10-5.20 (2H, m).

N,N′,N″-trigeranylcitric Triamide

¹H NMR(CDCl₃) δ=1.60 (9H, s), 1.66 (9H, s), 1.68 (9H, s), 1.98-2.08(12H, m), 3.76 (6H, t, J=6.3 Hz), 4.26 (4H, s), 5.07 (6H, t, J6.0 Hz),5.20 (6H, t, J=7.2 Hz).

N-geranylsuccinic Monoamide

¹H NMR(CDCl₃) δ=1.60 (3H, s), 1.70 (3H, s), 1.72 (3H, s), 1.92-2.15 (4H,m), 2.52 (2H, t, J=9.6 Hz), 2.70 (2H, t, J=9.6 Hz), 3.80-3.90 (2H, m),5.08 (1H, t, J=9.6 Hz), 5.18 (1H, t, J=6.0 Hz), 5.61 (1H, brs).

N,N′-digeranylsuccinic diamide, which was synthesized by a similarmethod, using N-geranylsuccenic monoamide as the starting material.

¹H NMR(CDCl₃) δ=1.60 (6H, s), 1.66 (6H, s), 1.69 (6H, s), 1.97-2.11 (4H,m), 2.53 (4H, s), 3.84 (4H, t, J=5.5 Hz), 5.07 (2H, t, J=4.9 Hz), 5.17(2H, t, J=5.5 Hz), 5.90 (2H, brs).

N-geranylfumaric Monoamide

¹H NMR(CDCl₃) δ=1.59 (3H, s), 1.67 (3H, s), 1.70 (3H, s), 1.94-2.16 (4H,m), 3.88-4.04 (2H, m), 5.06 (1H, t, J=7.2 Hz), 5.21 (1H, t, J=4.8 Hz),6.30 (1H, d, J=12.0 Hz), 6.46 (1H, d, J=12.0 Hz).

N,N′-digeranyl-2-Oxoglutaric Diamide

¹H NMR(CDCl₃) δ=1.60 (6H, s), 1.68 (12H, s), 1.94-2.13 (8H, m), 2.69(2H, t, J=6.3 Hz), 3.26 (2H, t, J=6.3 Hz), 3.81-4.04 (4H, m), 5.02-5.10(2H, m), 5.15-5.22 (2H, m).

N-geranyl-p-pyruvoaminobenzamide

¹H NMR(CDCl₃) δ=1.60 (3H, s), 1.68 (3H, s), 1.70 (3H, s), 2.03-2.11 (4H,m), 2.17 (3H, s), 3.95-4.04 (2H, m), 4.83 (1H, brs), 5.09 (1H, t, J=6.6Hz), 5.28 (1H, t, J=6.9 Hz), 5.94 (1H, brs), 6.64 (2H, d, J=8.7 Hz),7.60 (2H, d, J=8.7 Hz).

N-geranylretinamide

¹H NMR(CDCl₃) δ=1.03 (6H, s), 1.12-1.63 (6H, m), 1.60 (3H, s), 1.66 (3H,s), 1.68 (3H, s), 1.72 (3H, s), 1.87-1.93 (4H, m), 2.01 (3H, s), 2.37(3H, s), 3.82-3.92 (2H, m), 5.03-5.24 (2H, m), 5.80 (1H, s), 6.12-6.40(3H, m), 7.02 (1H, d, J=12.0 Hz), 7.07 (1H, d, J=12.0 Hz).

Synthesis Example 8 Synthesis of N-geranoylcysteine

To a THF (20 ml) solution containing geranic acid (1.68 g, 10 mmols),triethylamine (1.01 g, 10 mmols) was added and cooled to 0° C., intowhich a THF (5 ml) solution of isobutyl chloroformate (1.37 g, 10 mmols)was added dropwise, followed by 30 minutes' stirring at 0° C. To thereaction mixture a solution of cysteine (1.35 g, 10 mmols) as dissolvedin 1N sodium hydroxide (10 ml) was added, followed by an hour's stirringat 0° C. and further 4 hours' stirring at room temperature. Aftertermination of the reaction, 10 ml of 1N hydrochloric acid was added tothe reaction mixture and stirred for 10 minutes at room temperature.Then the reaction mixture was condensed with a rotary evaporator. To theresidue ethanol was added, and whereupon precipitated sodium chloridewas removed by filtration. The ethanol solution was once again condensedunder reduced pressure with the evaporator, and the residue wasseparated on silica gel column chromatography. Thus 0.556 g of the titlecompound was obtained from the hexane-acetone (2:1) distillate. Theyield was 19.5%:

¹H NMR(CDCl₃) δ=1.59 (6H, s), 1.68 (3H, s), 2.00-2.24 (4H, m), 2.60-2.77(1H, m), 3.00-3.30 (2H, m), 4.48-4.58 (1H, m), 5.00-5.13 (1H, m), 8.96(1H, s).

Synthesis Example 9 N-geranoyltyrosine

Synthesis Example 8 was repeated except that the cysteine was replacedwith tyrosine, to provide the title compound:

¹H NMR(CDCl₃) δ=1.55 (6H, s), 1.64 (3H, s), 1.96-2.00 (4H, m), 2.90-3.17(2H, m), 4.81-5.06 (3H, m), 6.40-7.21 (4H, m), 7.25 (1H, s).

Synthesis Example 10 Synthesis of Glutamic Digeranyldiamide

To a THF (20 ml) solution of N-benzyloxycarbonylglutamic acid (2.634 g,9.4 mmols), triethylamine (1.899 g, 18.8 mmols) was added and cooled to0° C. Into this mixture a THF (10 ml) solution of isobutyl chloroformate(2.566 g, 18.8 mmols) was added dropwise, followed by 30 minutes'stirring at 0° C. Then a THF (10 ml) solution of geranylamine (2.880 g,18.8 mmols) was added dropwise, followed by an hour's stirring at 0° C.and further 4 hours' stirring at room temperature. After termination ofthe reaction, 150 ml of chloroform was added to the system, and thechloroform solution was washed with water and dried over magnesiumsulfate. The organic solvent was removed with an evaporator and theresidue was separated on silica gel column chromatography. Thus 3.034 gof N-benzyloxycarbonylglutamic digeranyldiamide was obtained from thehexane-acetone (2:1) distillate. The yield was 58.6%.

Then the N-benzyloxycarbonylglutamic digeranyldiamide (3.034 g, 5.5mmols) was dissolved in methanol (20 ml), and to the same solution 20 mlof 1N sodium hydroxide was added, followed by 5 hours' stirring at roomtemperature. The reaction mixture was condensed with an evaporator, andthe residue was separated on silica gel column chromatography to providethe object compound from the hexane-ethanol (3:1) distillate. Becausethe as-obtained product was viscous and amorphous, ether was added tothe product for crystallization. Upon suction-filtering the system, 852mg of the object compound was obtained. The yield was 37.2%:

¹H NMR(CDCl₃) δ=1.58 (12H, s), 1.61 (6H, s), 1.75-2.12 (8H, m),2.32-2.53 (2H, m), 3.54-3.88 (7H, m), 4.88-5.21 (4H, m).

Synthesis Example 11

Repeating Synthesis Example 10 except that N-benzyloxycarbonylglutamicacid was replaced with tyrosine, N-acetyltyrosine, cysteine or serine,the following compounds, respectively, were obtained. WhereN-acetyltyrosine was used, the later deprotection operation was notconducted.

Tyrosine Geranylamide

¹H NMR(CDCl₃) δ=1.59 (3H, s), 1.67 (6H, s), 1.82-2.18 (4H, m), 2.99-3.09(2H, m), 3.74-3.78 (2H, m), 4.99-5.26 (3H, m), 7.17-7.43 (5H, m).

N-acetyltyrosine Geranylamide

¹H NMR(CDCl₃) δ=1.60 (3H, s), 1.68 (3H, s), 1.98 (3H, s), 2.00-2.11 (4H,m), 2.18 (3H, s), 2.90-3.00 (2H, m), 3.69-3.79 (2H, m), 4.59 (1H, dd,J=15.6, 9.6 Hz), 5.00-5.10 (2H, m), 6.70 (2H, d, J=7.8 Hz), 7.01 (2H, d,J=7.8 Hz), 7.27 (1H, s).

Cysteine Geranylamide

¹H NMR(CDCl₃) δ=1.58 (3H, s), 1.66 (3H, s), 1.67 (3H, s), 1.93-2.10 (4H,m), 2.83-3.16 (2H, m), 3.83-4.08 (3H, m), 5.03-5.19 (2H, m), 7.33 (1H,s).

Serine Geranylamide

¹H NMR(CDCl₃) δ=1.59 (3H, s), 1.68 (6H, s), 1.95-2.14 (4H, m), 3.80-3.95(2H, m), 4.34-4.47 (2H, m), 4.67 (1H, t, J=10.8 Hz), 5.06 (1H, t. J=6.0Hz), 5.17 (1H, t, J=6.0 Hz), 6.77 (2H, brs).

Synthesis Example 12 Synthesis of N-glucosylmevalonamide

Glucosamine hydrochloride (2.16 g, 10 mmols) was dissolved in 20 ml ofwater, and to the aqueous solution 10 ml of 1N sodium hydroxide andmevalolactone (1.30 g, 10 mmols) were added, followed by 5 hours'heating under stirring at 55° C. After termination of the reaction, thereaction mixture was condensed under reduced pressure. To the residue100 ml of methanol was added and whereupon separated precipitate wasfiltered off. The filtrate was condensed again with an evaporator andthe residue was separated on silica gel column chromatography, toprovide 1.45 g of the object product from the ethanol distillate. Theyield was 47%. Because the as-obtained product was a viscous oilysubstance, a minor amount of dichloromethane was added thereto to effectcrystallization. Upon suction filtering, 1.10 g of the title compoundwas obtained, which was strongly hygroscopic and its melting point couldnot be measured:

1H NMR (DMSO-d6) δ=1.00 (3H, s), 1.44-1.59 (2H, m), 2.47 (2H, s),2.96-3.74 (10H, m), 4.04-5.08 (3H, m).

Synthesis Example 13 Synthesis of N-galactosylmevalonamide

Synthesis Example 12 was repeated except that the glucosaminehydrochloride was replaced with galactosamine hydrochloride, to providethe title compound:

¹H NMR (DMSO-d6) δ=1.08 (3H, s), 1.51-1.61 (2H, m), 2.44 (2H, s),2.74-5.16 (13H, m).

Synthesis Example 14 Synthesis of N-fucosemevalonamide

Synthesis Example 12 was repeated except that the glucosaminehydrochloride was replaced with fucosamine hydrochloride, to provide thetitle compound:

¹H NMR (DMSO-d6) δ=1.06 (3H, s), 1.20 (3H, d, J=24.0 Hz), 1.54-1.62 (2H,m), 2.44 (2H, s), 2.74-5.15 (12H, m).

Formulation Example 1

Two (2) g of N-galactosylgeranamide was dissolved in 1 liter of waterfor injection at ambient temperature, isotonized with sodium chlorideand sealed into ampoules. One (1) ml of this injection contains 2 mg ofthe active ingredient.

Formulation Example 2

Two (2) g of N,N′-digeranylmalic diamide was dissolved in 1 liter ofwater for injection at ambient temperature, isotonized with sodiumchloride and sealed into ampoules. One (1) ml of this injection contains2 mg of the active ingredient.

Formulation Example 3

Two (2) g of N-glucosylmevalonamide was dissolved in 1 liter of waterfor injection at ambient temperature, isotonized with sodium chlorideand sealed into ampoules. One (1) ml of this injection contains 2 mg ofthe active ingredient.

1. A geranyl compound represented by the following formula (I-3):

wherein R³ stands for p-hydroxyphenyl or mercapto group.
 2. An agentcontaining, as an active ingredient, the geranyl compound of the formula(I-3) as claimed in claim
 1. 3. A pharmaceutical composition comprisingthe geranyl compound of the formula (I-3) as claimed in claim 1, and apharmaceutically acceptable carrier or diluent.
 4. A method of treatingsolid tumors of the liver comprising administering as antitumoricallyeffective amount of the geranyl compound of the formula (I-3) as claimedin claim 1 to a patient in need thereof.
 5. A method of making thepharmaceutical composition according to claim 3, which comprises mixingthe geranyl compound of the formula (I-3) with the pharmaceuticallyacceptable carrier or diluent.